Related Experiment Video
Updated: Apr 23, 2026
Bio-layer Interferometry for Measuring Kinetics of Protein-protein Interactions and Allosteric Ligand Effects
Published on: February 18, 2014
Kinetic titration series with biolayer interferometry
Daniel Frenzel1, Dieter Willbold2
1Forschungszentrum Jülich, ICS-6 Structural Biochemistry, Jülich, Germany.
Insights
This study demonstrates kinetic titration series for biolayer interferometry, enabling precise, real-time analysis of high-affinity protein interactions like immunoglobulin G and scFv IC16 with amyloid beta. This method enhances efficiency and accuracy in characterizing molecular binding events.
Area of Science:
- Biochemistry
- Molecular Biology
- Analytical Chemistry
Background:
- Biolayer interferometry (BLI) is a label-free optical technique for real-time analysis of biomolecular interactions.
- Characterizing high-affinity protein-ligand interactions quantitatively is crucial in drug discovery and diagnostics.
- Kinetic titration series are established for surface plasmon resonance (SPR) but their application in BLI requires validation.
Purpose of the Study:
- To demonstrate the straightforward application of kinetic titration series to biolayer interferometry for quantitative analysis of protein-ligand interactions.
- To characterize the binding kinetics of immunoglobulin G (IgG) with Protein G B1 and scFv IC16 with amyloid beta (1-42) using this method.
- To highlight the advantages of kinetic titration series in BLI, including circumvention of sensor variability, resource savings, and increased throughput.
Main Methods:
- Utilized biolayer interferometry for real-time interaction analysis.
- Employed kinetic titration series, involving sequential analyte injections over a concentration range on a single ligand-coated sensor chip.
- Characterized interactions between immunoglobulin G and Protein G B1, and between scFv IC16 and amyloid beta (1-42).
Main Results:
- Successfully applied kinetic titration series to biolayer interferometry for quantitative analysis of high-affinity protein-ligand interactions.
- Demonstrated the method's ability to accurately characterize binding kinetics for both IgG/Protein G B1 and scFv IC16/amyloid beta pairs.
- Validated that the BLI kinetic titration approach circumvents data evaluation issues from sensor differences.
Conclusions:
- Kinetic titration series are effectively transferable to biolayer interferometry, offering a robust method for real-time, quantitative interaction analysis.
- This approach significantly improves resource efficiency and throughput for screening numerous analyte/ligand combinations.
- The method provides a valuable tool for researchers studying high-affinity biomolecular interactions in various biological contexts.
Abstract:
Biolayer interferometry is a method to analyze protein interactions in real-time. In this study, we illustrate the usefulness to quantitatively analyze high affinity protein ligand interactions employing a kinetic titration series for characterizing the interactions between two pairs of interaction patterns, in particular immunoglobulin G and protein G B1 as well as scFv IC16 and amyloid beta (1-42). Kinetic titration series are commonly used in surface plasmon resonance and involve sequential injections of analyte over a desired concentration range on a single ligand coated sensor chip without waiting for complete dissociation between the injections. We show that applying this method to biolayer interferometry is straightforward and i) circumvents problems in data evaluation caused by unavoidable sensor differences, ii) saves resources and iii) increases throughput if screening a multitude of different analyte/ligand combinations.
Related Concept Videos
01:08Titrimetric Methods: Types and Commonly Used Strategies
02:18The Equilibrium Binding Constant and Binding Strength